LOCUS       sequence-374848-       13013 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  The construct was used for expression of gRNA and Cas9 for knocking 
            out of the ALDH1A3 gene..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13013)
  AUTHORS   Li J, Garavaglia S, Ye Z, Moretti A, Belyaeva OV, Beiser A, Ibrahim 
            M, Wilk A, McClellan S, Klyuyeva AV, Goggans KR, Kedishvili NY, 
            Salter EA, Wierzbicki A, Migaud ME, Mullett SJ, Yates NA, Camacho 
            CJ, Rizzi M, Sobol RW
  TITLE     A specific inhibitor of ALDH1A3 regulates retinoic acid biosynthesis
            in glioma stem cells.
  JOURNAL   Commun Biol. 2021 Dec 21;4(1):1420. doi: 10.1038/s42003-021-02949-7.
  PUBMED    34934174
REFERENCE   2  (bases 1 to 13013)
  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: "Commun 
            Biol."; date: "2021-12-21"; volume: "4(1):1420. doi"; pages: " 
            10.1038/s42003-021-02949-7"
COMMENT     Sequence Label: pLenti-v2-ALDH1A3gRNA1
FEATURES             Location/Qualifiers
     source          1..13013
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(115..134)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      928..1516
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1417..1436
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1670..1687
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    1804..1825
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(1840..1845,1846..1863,1864..1870)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1878..1894
                     /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)."
     primer_bind     1883..1905
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     1902..1918
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1902..1918
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    1967..2088
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1978..1997)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2032..2051
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(2218..2592)
                     /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"
     promoter        complement(2611..2658)
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     promoter        complement(2706..3035)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2720..2855
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(2774..2793)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     3020..3040
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      complement(3049..3477)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3159..3180)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3371..3390
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     polyA_signal    3523..3747
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     3736..3753
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     LTR             3779..4012
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     misc_feature    4084..4672
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             4190..4201
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     4599..4619
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4688..5284)
                     /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     complement(4771..4791)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     CDS             complement(5285..5341)
                     /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             complement(5351..5374)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             complement(5375..5422)
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             complement(5423..9526)
                     /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             complement(5423..9526)
                     /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"
     regulatory      9523..9532
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(9551..9762)
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     misc_RNA        complement(9824..9899)
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        complement(9921..10169)
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     complement(9979..9998)
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     primer_bind     complement(10149..10169)
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     misc_feature    10220..10337
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     CDS             complement(10635..10679)
                     /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    10864..11097
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     misc_feature    11590..11715
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             11762..11942
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     promoter        complement(11960..12158)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(11985..12005)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        12160..12539
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     12711..12730
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        12805..12909
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(12910..12978,12979..13013,1..757)
                     /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"
ORIGIN
        1 agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct
       61 tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg
      121 tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta
      181 ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat
      241 gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt
      301 acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga
      361 tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga
      421 gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga
      481 actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc
      541 aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc
      601 cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg
      661 tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat
      721 cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata
      781 tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct
      841 ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga
      901 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
      961 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
     1021 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct
     1081 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     1141 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     1201 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     1261 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
     1321 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
     1381 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
     1441 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
     1501 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
     1561 gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac
     1621 cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt
     1681 gagcgaggaa gcggaagagc gcccaatacg caaaccgcct ctccccgcgc gttggccgat
     1741 tcattaatgc agctggcacg acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc
     1801 aattaatgtg agttagctca ctcattaggc accccaggct ttacacttta tgcttccggc
     1861 tcgtatgttg tgtggaattg tgagcggata acaatttcac acaggaaaca gctatgacca
     1921 tgattacgcc aagctctagc tagaggtcga cggtatacag acatgataag atacattgat
     1981 gagtttggac aaaccacaac tagaatgcag tgaaaaaaat gctttatttg tgaaatttgt
     2041 gatgctattg ctttatttgt aaccattata agctgcaata aacaagttgg ggtgggcgaa
     2101 gaactccagc atgagatccc cgcgctggag gatcatccag ccggcgtccc ggaaaacgat
     2161 tccgaagccc aacctttcat agaaggcggc ggtggaatcg aaatctcgta gcacgtgtca
     2221 gtcctgctcc tcggccacga agtgcacgca gttgccggcc gggtcgcgca gggcgaactc
     2281 ccgcccccac ggctgctcgc cgatctcggt catggccggc ccggaggcgt cccggaagtt
     2341 cgtggacacg acctccgacc actcggcgta cagctcgtcc aggccgcgca cccacaccca
     2401 ggccagggtg ttgtccggca ccacctggtc ctggaccgcg ctgatgaaca gggtcacgtc
     2461 gtcccggacc acaccggcga agtcgtcctc cacgaagtcc cgggagaacc cgagccggtc
     2521 ggtccagaac tcgaccgctc cggcgacgtc gcgcgcggtg agcaccggaa cggcactggt
     2581 caacttggcc atggtttagt tcctcacctt gtcgtattat actatgccga tatactatgc
     2641 cgatgattaa ttgtcaacac gtgctgatca gatccgaaaa tggatataca agctcccggg
     2701 agctttttgc aaaagcctag gcctccaaaa aagcctcctc actacttctg gaatagctca
     2761 gaggcagagg cggcctcggc ctctgcataa ataaaaaaaa ttagtcagcc atggggcgga
     2821 gaatgggcgg aactgggcgg agttaggggc gggatgggcg gagttagggg cgggactatg
     2881 gttgctgact aattgagatg catgctttgc atacttctgc ctgctgggga gcctggggac
     2941 tttccacacc tggttgctga ctaattgaga tgcatgcttt gcatacttct gcctgctggg
     3001 gagcctgggg actttccaca ccctaactga cacacattcc acagaattaa ttcgcgttaa
     3061 atttttgtta aatcagctca ttttttaacc aataggccga aatcggcaaa atcccttata
     3121 aatcaaaaga atagaccgag atagggttga gtgttgttcc agtttggaac aagagtccac
     3181 tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc
     3241 cactacgtga accatcaccc taatcaagtt ttttggggtc gaggtgccgt aaagcactaa
     3301 atcggaaccc taaagggagc ccccgattta gagcttgacg gggaaagccg gcgaacgtgg
     3361 cgagaaagga agggaagaaa gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg
     3421 tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc gccgctacag ggcgcgtggg
     3481 gataccccct agagccccag ctggttcttt ccgcctcaga agccatagag cccaccgcat
     3541 ccccagcatg cctgctattg tcttcccaat cctccccctt gctgtcctgc cccaccccac
     3601 cccccagaat agaatgacac ctactcagac aatgcgatgc aatttcctca ttttattagg
     3661 aaaggacagt gggagtggca ccttccaggg tcaaggaagg cacgggggag gggcaaacaa
     3721 cagatggctg gcaactagaa ggcacagtcg aggctgatca gcgggtttaa acgggccctg
     3781 ctagagattt tccacactga ctaaaagggt ctgagggatc tctagttacc agagtcacac
     3841 aacagacggg cacacactac ttgaagcact caaggcaagc tttattgagg cttaagcagt
     3901 gggttcccta gttagccaga gagctcccag gctcagatct ggtctaacca gagagaccca
     3961 gtacaagcaa aaagcagatc ttgtcttcgt tgggagtgaa ttagcccttc cagtcccccc
     4021 ttttctttta aaaagtggct aagatctaca gctgccttgt aagtcattgg tcttaaagtc
     4081 gacgcgggga ggcggcccaa agggagatcc gactcgtctg agggcgaagg cgaagacgcg
     4141 gaagaggccg cagagccggc agcaggccgc gggaaggaag gtccgctgga ttgagggccg
     4201 aagggacgta gcagaaggac gtcccgcgca gaatccaggt ggcaacacag gcgagcagcc
     4261 aaggaaagga cgatgatttc cccgacaaca ccacggaatt gtcagtgccc aacagccgag
     4321 cccctgtcca gcagcgggca aggcaggcgg cgatgagttc cgccgtggca atagggaggg
     4381 ggaaagcgaa agtcccggaa aggagctgac aggtggtggc aatgccccaa ccagtggggg
     4441 ttgcgtcagc aaacacagtg cacaccacgc cacgttgcct gacaacgggc cacaactcct
     4501 cataaagaga cagcaaccag gatttataca aggaggagaa aatgaaagcc atacgggaag
     4561 caatagcatg atacaaaggc attaaagcag cgtatccaca tagcgtaaaa ggagcaacat
     4621 agttaagaat accagtcaat ctttcacaaa ttttgtaatc cagaggttga ttgtcgactt
     4681 aacgcgttca ggcaccgggc ttgcgggtca tgcaccaggt gcgcggtcct tcgggcacct
     4741 cgacgtcggc ggtgacggtg aagccgagcc gctcgtagaa ggggaggttg cggggcgcgg
     4801 aggtctccag gaaggcgggc accccggcgc gctcggccgc ctccactccg gggagcacga
     4861 cggcgctgcc cagacccttg ccctggtggt cgggcgagac tccgacggtg gccaggaacc
     4921 acgcgggctc cttgggccgg tgcggcgcca ggaggccttc catctgttgc tgcgcggcca
     4981 gccgggaacc gctcaactcg gccatgcgcg ggccgatctc ggcgaacacc gcccccgctt
     5041 cgacgctctc cggcgtggtc cagaccgcca ccgcggcgcc gtcgtccgcg acccacacct
     5101 tgccgatgtc gagcccgacg cgcgtgagga agagttcttg cagctcggtg acccgctcga
     5161 tgtggcggtc cggatcgacg gtgtggcgcg tggcggggta gtcggcgaac gcggcggcga
     5221 gggtgcgtac ggccctgggg acgtcgtcgc gggtggcgag gcgcaccgtg ggcttgtact
     5281 cggtcggtcc aggattctct tcgacatctc cggcttgttt cagcagagag aagtttgttg
     5341 cgccggatcc cttatcgtca tcgtctttgt aatctttctt cttcttagcc tgtccagcct
     5401 tctttgtggc ggcaggtcgc ttgtcgcctc ccagctgaga caggtcgatc cgtgtctcgt
     5461 acaggccggt gatgctctgg tggatcaggg tggcgtccag cacctctttg gtgctggtgt
     5521 acctcttccg gtcgatggtg gtgtcaaagt acttgaaggc ggcaggggct cccagattgg
     5581 tcagggtaaa caggtggatg atattctcgg cctgctctct gatgggctta tcccggtgct
     5641 tgttgtaggc ggacagcact ttgtccagat tagcgtcggc caggatcact ctcttggaga
     5701 actcgctgat ctgctcgatg atctcgtcca ggtagtgctt gtgctgttcc acaaacagct
     5761 gtttctgctc attatcctcg ggggagccct tcagcttctc atagtggctg gccaggtaca
     5821 ggaagttcac atatttggag ggcagggcca gttcgtttcc cttctgcagt tcgccggcag
     5881 aggccagcat tctcttccgg ccgttttcca gctcgaacag ggagtactta ggcagcttga
     5941 tgatcaggtc ctttttcact tctttgtagc ccttggcttc cagaaagtcg atgggattct
     6001 tctcgaagct gcttctttcc atgatggtga tccccagcag ctctttcaca ctcttcagtt
     6061 tcttggactt gcccttttcc actttggcca ccaccagcac agaataggcc acggtggggc
     6121 tgtcgaagcc gccgtacttc ttagggtccc agtccttctt tctggcgatc agcttatcgc
     6181 tgttcctctt gggcaggata gactctttgc tgaagccgcc tgtctgcacc tcggtctttt
     6241 tcacgatatt cacttggggc atgctcagca ctttccgcac ggtggcaaaa tcccggccct
     6301 tatcccacac gatctccccg gtttcgccgt ttgtctcgat cagaggccgc ttccggatct
     6361 cgccgttggc cagggtaatc tcggtcttga aaaagttcat gatgttgctg tagaagaagt
     6421 acttggcggt agccttgccg atttcctgct cgctcttggc gatcatcttc cgcacgtcgt
     6481 acaccttgta gtcgccgtac acgaactcgc tttccagctt agggtacttt ttgatcaggg
     6541 cggttcccac gacggcgttc aggtaggcgt cgtgggcgtg gtggtagttg ttgatctcgc
     6601 gcactttgta aaactggaaa tccttccgga aatcggacac cagcttggac ttcagggtga
     6661 tcactttcac ttcccggatc agcttgtcat tctcgtcgta cttagtgttc atccgggagt
     6721 ccaggatctg tgccacgtgc tttgtgatct gccgggtttc caccagctgt ctcttgatga
     6781 agccggcctt atccagttcg ctcaggccgc ctctctcggc cttggtcaga ttgtcgaact
     6841 ttctctgggt aatcagcttg gcgttcagca gctgccgcca gtagttcttc atcttcttca
     6901 cgacctcttc ggagggcacg ttgtcgctct tgccccggtt cttgtcgctt ctggtcagca
     6961 ccttgttgtc gatggagtcg tccttcagaa agctctgagg cacgatatgg tccacatcgt
     7021 agtcggacag ccggttgatg tccagttcct ggtccacgta catatcccgc ccattctgca
     7081 ggtagtacag gtacagcttc tcgttctgca gctgggtgtt ttccacgggg tgttctttca
     7141 ggatctggct gcccagctct ttgatgccct cttcgatccg cttcattctc tcgcggctgt
     7201 tcttctgtcc cttctgggtg gtctggttct ctctggccat ttcgatcacg atgttctcgg
     7261 gcttgtgccg gcccatcact ttcacgagct cgtccaccac cttcactgtc tgcaggatgc
     7321 ccttcttaat ggcggggctg ccggccagat tggcaatgtg ctcgtgcagg ctatcgccct
     7381 ggccggacac ctgggctttc tggatgtcct ctttaaaggt caggctgtcg tcgtggatca
     7441 gctgcatgaa gtttctgttg gcgaagccgt cggacttcag gaaatccagg attgtcttgc
     7501 cggactgctt gtcccggatg ccgttgatca gcttccggct cagcctgccc cagccggtgt
     7561 atctccgccg cttcagctgc ttcatcactt tgtcgtcgaa caggtgggca taggttttca
     7621 gccgttcctc gatcatctct ctgtcctcaa acagtgtcag ggtcagcacg atatcttcca
     7681 gaatgtcctc gttttcctca ttgtccagga agtccttgtc cttgataatt ttcagcagat
     7741 cgtggtatgt gcccagggag gcgttgaacc gatcttccac gccggagatt tccacggagt
     7801 cgaagcactc gattttcttg aagtagtcct ctttcagctg cttcacggtc actttccggt
     7861 tggtcttgaa cagcaggtcc acgatggcct ttttctgctc gccgctcagg aaggcgggct
     7921 ttctcattcc ctcggtcacg tatttcactt tggtcagctc gttatacacg gtgaagtact
     7981 cgtacagcag gctgtgcttg ggcagcacct tctcgttggg caggttctta tcgaagttgg
     8041 tcatccgctc gatgaagctc tgggcggaag cgcccttgtc caccacttcc tcgaagttcc
     8101 agggggtgat ggtttcctcg ctctttctgg tcatccaggc gaatctgctg tttcccctgg
     8161 ccagagggcc cacgtagtag gggatgcgga aggtcaggat cttctcgatc ttttcccggt
     8221 tgtccttcag gaatgggtaa aaatcttcct gccgccgcag aatggcgtgc agctctccca
     8281 ggtggatctg gtgggggatg ctgccgttgt cgaaggtccg ctgcttccgc agcaggtcct
     8341 ctctgttcag cttcacgagc agttcctcgg tgccgtccat cttttccagg atgggcttga
     8401 tgaacttgta gaactcttcc tggctggctc cgccgtcaat gtagccggcg tagccgttct
     8461 tgctctggtc gaagaaaatc tctttgtact tctcaggcag ctgctgccgc acgagagctt
     8521 tcagcagggt caggtcctgg tggtgctcgt cgtatctctt gatcatagag gcgctcaggg
     8581 gggccttggt gatctcggtg ttcactctca ggatgtcgct cagcaggatg gcgtcggaca
     8641 ggttcttggc ggccagaaac aggtcggcgt actggtcgcc gatctgggcc agcaggttgt
     8701 ccaggtcgtc gtcgtaggtg tccttgctca gctgcagttt ggcatcctcg gccaggtcga
     8761 agttgctctt gaagttgggg gtcaggccca ggctcagggc aatcaggttt ccgaacaggc
     8821 cattcttctt ctcgccgggc agctgggcga tcagattttc cagccgtctg ctcttgctca
     8881 gtctggcaga caggatggcc ttggcgtcca cgccgctggc gttgatgggg ttttcctcga
     8941 acagctggtt gtaggtctgc accagctgga tgaacagctt gtccacgtcg ctgttgtcgg
     9001 ggttcaggtc gccctcgatc aggaagtggc cccggaactt gatcatgtgg gccagggcca
     9061 gatagatcag ccgcaggtcg gccttgtcgg tgctgtccac cagtttcttt ctcaggtggt
     9121 agatggtggg gtacttctcg tggtaggcca cctcgtccac gatgttgccg aagatggggt
     9181 gccgctcgtg cttcttatcc tcttccacca ggaaggactc ttccagtctg tggaagaagc
     9241 tgtcgtccac cttggccatc tcgttgctga agatctcttg cagatagcag atccggttct
     9301 tccgtctggt gtatcttctt ctggcggttc tcttcagccg ggtggcctcg gctgtttcgc
     9361 cgctgtcgaa cagcagggct ccgatcaggt tcttcttgat gctgtgccgg tcggtgttgc
     9421 ccagcacctt gaatttcttg ctgggcacct tgtactcgtc ggtgatcacg gcccagccca
     9481 cagagttggt gccgatgtcc aggccgatgc tgtacttctt gtccatggtg gcagcgctct
     9541 agaaccggtc ctgtgttctg gcggcaaacc cgttgcgaaa aagaacgttc acggcgacta
     9601 ctgcacttat atacggttct cccccaccct cgggaaaaag gcggagccag tacacgacat
     9661 cactttccca gtttaccccg cgccaccttc tctaggcacc ggatcaattg ccgacccctc
     9721 cccccaactt ctcggggact gtgggcgatg tgcgctctgc ccactgacgg gcaccggagc
     9781 caattcccac tcctttcaag acctagctag cgaattcaaa aaagcaccga ctcggtgcca
     9841 ctttttcaag ttgataacgg actagcctta ttttaacttg ctatttctag ctctaaaact
     9901 ggaaaacggg cagccggacc ggtgtttcgt cctttccaca agatatataa agccaagaaa
     9961 tcgaaatact ttcaagttac ggtaagcata tgatagtcca ttttaaaaca taattttaaa
    10021 actgcaaact acccaagaaa ttattacttt ctacgtcacg tattttgtac taatatcttt
    10081 gtgtttacag tcaaattaat tctaattatc tctctaacag ccttgtatcg tatatgcaaa
    10141 tatgaaggaa tcatgggaaa taggccctcg gtaccttaat taaccaaact ggatctctgc
    10201 tgtccctgta ataaacccga aaattttgaa tttttgtaat ttgtttttgt aattctttag
    10261 tttgtatgtc tgttgctatt atgtctacta ttctttcccc tgcactgtac cccccaatcc
    10321 ccccttttct tttaaaattg tggatgaata ctgccatttg tctgcagaat tggcgcacgc
    10381 agtgccgatc cgttcactaa tcgaatggat ctgtctctgt ctctctctcc accttcttct
    10441 tctattcctt cgggcctgtc gggtcccctc ggggttggga ggtgggtctg aaacgataat
    10501 ggtgaatatc cctgcctaac tctattcact atagaaagta cagcaaaaac tattcttaaa
    10561 cctaccaagc ctcctactat cattatgaat aattttatat accacagcca atttgttatg
    10621 ttaaaccaat tccacaaact tgcccattta tctaattcca ataattcttg ttcattcttt
    10681 tcttgctggt tttgcgattc ttcaattaag gagtgtatta agcttgtgta attgttaatt
    10741 tctctgtccc actccatcca ggtcgtgtga ttccaaatct gttccagaga tttattactc
    10801 caactagcat tccaaggcac agcagtggtg caaatgagtt ttccagagca accccaaatc
    10861 cccaggagct gttgatcctt taggtatctt tccacagcca ggattcttgc ctggagctgc
    10921 ttgatgcccc agactgtgag ttgcaacaga tgctgttgcg cctcaatagc cctcagcaaa
    10981 ttgttctgct gctgcactat accagacaat aattgtctgg cctgtaccgt cagcgtcatt
    11041 gacgctgcgc ccatagtgct tcctgctgct cccaagaacc caaggaacaa agctcctatt
    11101 cccactgctc ttttttctct ctgcaccact cttctctttg ccttggtggg tgctactcct
    11161 aatggttcaa tttttactac tttatattta tataattcac ttctccaatt gtccctcata
    11221 tctcctcctc caggtctgaa gatcagcggc cgcttgctgt gcggtggtct tacttttgtt
    11281 ttgctcttcc tctatcttgt ctaaagcttc cttggtgtct tttatctcta tcctttgatg
    11341 cacacaatag agggttgcta ctgtattata taatgatcta agttcttctg atcctgtctg
    11401 aagggatggt tgtagctgtc ccagtatttg tctacagcct tctgatgttt ctaacaggcc
    11461 aggattaact gcgaatcgtt ctagctccct gcttgcccat actatatgtt ttaatttata
    11521 ttttttcttt ccccctggcc ttaaccgaat tttttcccat cgcgatctaa ttctcccccg
    11581 cttaatactg acgctctcgc acccatctct ctccttctag cctccgctag tcaaaatttt
    11641 tggcgtactc accagtcgcc gcccctcgcc tcttgccgtg cgcgcttcag caagccgagt
    11701 cctgcgtcga gagagctcct ctggtttccc tttcgctttc aagtccctgt tcgggcgcca
    11761 ctgctagaga ttttccacac tgactaaaag ggtctgaggg atctctagtt accagagtca
    11821 cacaacagac gggcacacac tacttgaagc actcaaggca agctttattg aggcttaagc
    11881 agtgggttcc ctagttagcc agagagctcc caggctcaga tctggtctaa ccagagagac
    11941 ccagtacagg caaaacgcgc tgcttatata gacctcccac cgtacacgcc taccgcccat
    12001 ttgcgtcaat ggggcggagt tgttacgaca ttttggaaag tcccgttgat tttggtgcca
    12061 aaacaaactc ccattgacgt caatggggtg gagacttgga aatccccgtg agtcaaaccg
    12121 ctatccacgc ccattgatgt actgccaaaa ccgcatcacc atggtaatag cgatgactaa
    12181 tacgtagatg tactgccaag taggaaagtc ccataaggtc atgtactggg cataatgcca
    12241 ggcgggccat ttaccgtcat tgacgtcaat agggggcgta cttggcatat gatacacttg
    12301 atgtactgcc aagtgggcag tttaccgtaa atactccacc cattgacgtc aatggaaagt
    12361 ccctattggc gttactatgg gaacatacgt cattattgac gtcaatgggc gggggtcgtt
    12421 gggcggtcag ccaggcgggc catttaccgt aagttatgta acgcggaact ccatatatgg
    12481 gctatgaact aatgaccccg taattgatta ctattaataa ctagtcaata atcaatgtca
    12541 acgcgtatat ctggcccgta catcgcgaag cagcgcaaaa cgcctaaccc taagcagatt
    12601 cttcatgcaa ttgtcggtca agccttgcct tgttgtagct taaattttgc tcgcgcacta
    12661 ctcagcgacc tccaacacac aagcagggag cagatactgg cttaactatg cggcatcaga
    12721 gcagattgta ctgagagtgc accatagggg atcgggagat ctcccgatcc gtcgacgtca
    12781 ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat
    12841 tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa
    12901 aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt
    12961 tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc tga
//
