LOCUS       sequence-332995-       13013 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Disrupt ACO1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13013)
  AUTHORS   Terzi EM, Sviderskiy VO, Alvarez SW, Whiten GC, Possemato R
  TITLE     Iron-sulfur cluster deficiency can be sensed by IRP2 and regulates 
            iron homeostasis and sensitivity to ferroptosis independent of IRP1 
            and FBXL5.
  JOURNAL   Sci Adv. 2021 May 26;7(22). pii: 7/22/eabg4302. doi: 
            10.1126/sciadv.abg4302. Print 2021 May.
  PUBMED    34039609
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: "Sci Adv."; 
            date: "2021-05-26"; volume: "7(22). pii: 7/22/eabg4302. doi"; pages:
            " 10.1126/sciadv.abg4302. Print 2021 May"
COMMENT     Sequence Label: pLentiCRISPR V2 sgACO1_1
FEATURES             Location/Qualifiers
     source          1..13013
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(164..183)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        355..734
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        736..934
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     889..909
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             952..1132
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    1179..1304
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1797..2030
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2215..2259
                     /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    2557..2674
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2725..2973
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     2725..2745
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     2896..2915
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        2995..3070
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        3132..3343
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     regulatory      3362..3371
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3368..7471
                     /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             3368..7471
                     /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             7472..7519
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             7520..7543
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             7553..7609
                     /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             7610..8206
                     /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     8103..8123
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    8222..8810
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(8275..8295)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(8693..8704)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             8882..9115
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(9141..9158)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    9147..9371
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      9417..9845
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(9504..9523)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     9714..9735
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(9854..9874)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        9859..10188
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      10039..10174
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     10101..10120
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        10236..10283
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             10302..10676
                     /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    10806..10927
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(10843..10862)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     10897..10916
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(10976..10992)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(10976..10992)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(10989..11011)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    11000..11016
                     /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(11024..11030,11031..11048,11049..11054))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    11069..11090
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(11207..11224)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(11378..11966)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(11458..11477)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(12137..12928,12929..12997))
                     /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     12760..12779
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(join(12998..13013,1..89))
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt
       61 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc
      121 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
      181 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      241 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      301 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      361 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      421 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      481 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      541 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      601 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      661 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      721 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      781 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      841 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      901 gtaggcgtgt acggtgggag gtctatataa gcagcgcgtt ttgcctgtac tgggtctctc
      961 tggttagacc agatctgagc ctgggagctc tctggctaac tagggaaccc actgcttaag
     1021 cctcaataaa gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct
     1081 ggtaactaga gatccctcag acccttttag tcagtgtgga aaatctctag cagtggcgcc
     1141 cgaacaggga cttgaaagcg aaagggaaac cagaggagct ctctcgacgc aggactcggc
     1201 ttgctgaagc gcgcacggca agaggcgagg ggcggcgact ggtgagtacg ccaaaaattt
     1261 tgactagcgg aggctagaag gagagagatg ggtgcgagag cgtcagtatt aagcggggga
     1321 gaattagatc gcgatgggaa aaaattcggt taaggccagg gggaaagaaa aaatataaat
     1381 taaaacatat agtatgggca agcagggagc tagaacgatt cgcagttaat cctggcctgt
     1441 tagaaacatc agaaggctgt agacaaatac tgggacagct acaaccatcc cttcagacag
     1501 gatcagaaga acttagatca ttatataata cagtagcaac cctctattgt gtgcatcaaa
     1561 ggatagagat aaaagacacc aaggaagctt tagacaagat agaggaagag caaaacaaaa
     1621 gtaagaccac cgcacagcaa gcggccgctg atcttcagac ctggaggagg agatatgagg
     1681 gacaattgga gaagtgaatt atataaatat aaagtagtaa aaattgaacc attaggagta
     1741 gcacccacca aggcaaagag aagagtggtg cagagagaaa aaagagcagt gggaatagga
     1801 gctttgttcc ttgggttctt gggagcagca ggaagcacta tgggcgcagc gtcaatgacg
     1861 ctgacggtac aggccagaca attattgtct ggtatagtgc agcagcagaa caatttgctg
     1921 agggctattg aggcgcaaca gcatctgttg caactcacag tctggggcat caagcagctc
     1981 caggcaagaa tcctggctgt ggaaagatac ctaaaggatc aacagctcct ggggatttgg
     2041 ggttgctctg gaaaactcat ttgcaccact gctgtgcctt ggaatgctag ttggagtaat
     2101 aaatctctgg aacagatttg gaatcacacg acctggatgg agtgggacag agaaattaac
     2161 aattacacaa gcttaataca ctccttaatt gaagaatcgc aaaaccagca agaaaagaat
     2221 gaacaagaat tattggaatt agataaatgg gcaagtttgt ggaattggtt taacataaca
     2281 aattggctgt ggtatataaa attattcata atgatagtag gaggcttggt aggtttaaga
     2341 atagtttttg ctgtactttc tatagtgaat agagttaggc agggatattc accattatcg
     2401 tttcagaccc acctcccaac cccgagggga cccgacaggc ccgaaggaat agaagaagaa
     2461 ggtggagaga gagacagaga cagatccatt cgattagtga acggatcggc actgcgtgcg
     2521 ccaattctgc agacaaatgg cagtattcat ccacaatttt aaaagaaaag gggggattgg
     2581 ggggtacagt gcaggggaaa gaatagtaga cataatagca acagacatac aaactaaaga
     2641 attacaaaaa caaattacaa aaattcaaaa ttttcgggtt tattacaggg acagcagaga
     2701 tccagtttgg ttaattaagg taccgagggc ctatttccca tgattccttc atatttgcat
     2761 atacgataca aggctgttag agagataatt agaattaatt tgactgtaaa cacaaagata
     2821 ttagtacaaa atacgtgacg tagaaagtaa taatttcttg ggtagtttgc agttttaaaa
     2881 ttatgtttta aaatggacta tcatatgctt accgtaactt gaaagtattt cgatttcttg
     2941 gctttatata tcttgtggaa aggacgaaac accgggtgga tgattcctga gccagtttta
     3001 gagctagaaa tagcaagtta aaataaggct agtccgttat caacttgaaa aagtggcacc
     3061 gagtcggtgc ttttttgaat tcgctagcta ggtcttgaaa ggagtgggaa ttggctccgg
     3121 tgcccgtcag tgggcagagc gcacatcgcc cacagtcccc gagaagttgg ggggaggggt
     3181 cggcaattga tccggtgcct agagaaggtg gcgcggggta aactgggaaa gtgatgtcgt
     3241 gtactggctc cgcctttttc ccgagggtgg gggagaaccg tatataagtg cagtagtcgc
     3301 cgtgaacgtt ctttttcgca acgggtttgc cgccagaaca caggaccggt tctagagcgc
     3361 tgccaccatg gacaagaagt acagcatcgg cctggacatc ggcaccaact ctgtgggctg
     3421 ggccgtgatc accgacgagt acaaggtgcc cagcaagaaa ttcaaggtgc tgggcaacac
     3481 cgaccggcac agcatcaaga agaacctgat cggagccctg ctgttcgaca gcggcgaaac
     3541 agccgaggcc acccggctga agagaaccgc cagaagaaga tacaccagac ggaagaaccg
     3601 gatctgctat ctgcaagaga tcttcagcaa cgagatggcc aaggtggacg acagcttctt
     3661 ccacagactg gaagagtcct tcctggtgga agaggataag aagcacgagc ggcaccccat
     3721 cttcggcaac atcgtggacg aggtggccta ccacgagaag taccccacca tctaccacct
     3781 gagaaagaaa ctggtggaca gcaccgacaa ggccgacctg cggctgatct atctggccct
     3841 ggcccacatg atcaagttcc ggggccactt cctgatcgag ggcgacctga accccgacaa
     3901 cagcgacgtg gacaagctgt tcatccagct ggtgcagacc tacaaccagc tgttcgagga
     3961 aaaccccatc aacgccagcg gcgtggacgc caaggccatc ctgtctgcca gactgagcaa
     4021 gagcagacgg ctggaaaatc tgatcgccca gctgcccggc gagaagaaga atggcctgtt
     4081 cggaaacctg attgccctga gcctgggcct gacccccaac ttcaagagca acttcgacct
     4141 ggccgaggat gccaaactgc agctgagcaa ggacacctac gacgacgacc tggacaacct
     4201 gctggcccag atcggcgacc agtacgccga cctgtttctg gccgccaaga acctgtccga
     4261 cgccatcctg ctgagcgaca tcctgagagt gaacaccgag atcaccaagg cccccctgag
     4321 cgcctctatg atcaagagat acgacgagca ccaccaggac ctgaccctgc tgaaagctct
     4381 cgtgcggcag cagctgcctg agaagtacaa agagattttc ttcgaccaga gcaagaacgg
     4441 ctacgccggc tacattgacg gcggagccag ccaggaagag ttctacaagt tcatcaagcc
     4501 catcctggaa aagatggacg gcaccgagga actgctcgtg aagctgaaca gagaggacct
     4561 gctgcggaag cagcggacct tcgacaacgg cagcatcccc caccagatcc acctgggaga
     4621 gctgcacgcc attctgcggc ggcaggaaga tttttaccca ttcctgaagg acaaccggga
     4681 aaagatcgag aagatcctga ccttccgcat cccctactac gtgggccctc tggccagggg
     4741 aaacagcaga ttcgcctgga tgaccagaaa gagcgaggaa accatcaccc cctggaactt
     4801 cgaggaagtg gtggacaagg gcgcttccgc ccagagcttc atcgagcgga tgaccaactt
     4861 cgataagaac ctgcccaacg agaaggtgct gcccaagcac agcctgctgt acgagtactt
     4921 caccgtgtat aacgagctga ccaaagtgaa atacgtgacc gagggaatga gaaagcccgc
     4981 cttcctgagc ggcgagcaga aaaaggccat cgtggacctg ctgttcaaga ccaaccggaa
     5041 agtgaccgtg aagcagctga aagaggacta cttcaagaaa atcgagtgct tcgactccgt
     5101 ggaaatctcc ggcgtggaag atcggttcaa cgcctccctg ggcacatacc acgatctgct
     5161 gaaaattatc aaggacaagg acttcctgga caatgaggaa aacgaggaca ttctggaaga
     5221 tatcgtgctg accctgacac tgtttgagga cagagagatg atcgaggaac ggctgaaaac
     5281 ctatgcccac ctgttcgacg acaaagtgat gaagcagctg aagcggcgga gatacaccgg
     5341 ctggggcagg ctgagccgga agctgatcaa cggcatccgg gacaagcagt ccggcaagac
     5401 aatcctggat ttcctgaagt ccgacggctt cgccaacaga aacttcatgc agctgatcca
     5461 cgacgacagc ctgaccttta aagaggacat ccagaaagcc caggtgtccg gccagggcga
     5521 tagcctgcac gagcacattg ccaatctggc cggcagcccc gccattaaga agggcatcct
     5581 gcagacagtg aaggtggtgg acgagctcgt gaaagtgatg ggccggcaca agcccgagaa
     5641 catcgtgatc gaaatggcca gagagaacca gaccacccag aagggacaga agaacagccg
     5701 cgagagaatg aagcggatcg aagagggcat caaagagctg ggcagccaga tcctgaaaga
     5761 acaccccgtg gaaaacaccc agctgcagaa cgagaagctg tacctgtact acctgcagaa
     5821 tgggcgggat atgtacgtgg accaggaact ggacatcaac cggctgtccg actacgatgt
     5881 ggaccatatc gtgcctcaga gctttctgaa ggacgactcc atcgacaaca aggtgctgac
     5941 cagaagcgac aagaaccggg gcaagagcga caacgtgccc tccgaagagg tcgtgaagaa
     6001 gatgaagaac tactggcggc agctgctgaa cgccaagctg attacccaga gaaagttcga
     6061 caatctgacc aaggccgaga gaggcggcct gagcgaactg gataaggccg gcttcatcaa
     6121 gagacagctg gtggaaaccc ggcagatcac aaagcacgtg gcacagatcc tggactcccg
     6181 gatgaacact aagtacgacg agaatgacaa gctgatccgg gaagtgaaag tgatcaccct
     6241 gaagtccaag ctggtgtccg atttccggaa ggatttccag ttttacaaag tgcgcgagat
     6301 caacaactac caccacgccc acgacgccta cctgaacgcc gtcgtgggaa ccgccctgat
     6361 caaaaagtac cctaagctgg aaagcgagtt cgtgtacggc gactacaagg tgtacgacgt
     6421 gcggaagatg atcgccaaga gcgagcagga aatcggcaag gctaccgcca agtacttctt
     6481 ctacagcaac atcatgaact ttttcaagac cgagattacc ctggccaacg gcgagatccg
     6541 gaagcggcct ctgatcgaga caaacggcga aaccggggag atcgtgtggg ataagggccg
     6601 ggattttgcc accgtgcgga aagtgctgag catgccccaa gtgaatatcg tgaaaaagac
     6661 cgaggtgcag acaggcggct tcagcaaaga gtctatcctg cccaagagga acagcgataa
     6721 gctgatcgcc agaaagaagg actgggaccc taagaagtac ggcggcttcg acagccccac
     6781 cgtggcctat tctgtgctgg tggtggccaa agtggaaaag ggcaagtcca agaaactgaa
     6841 gagtgtgaaa gagctgctgg ggatcaccat catggaaaga agcagcttcg agaagaatcc
     6901 catcgacttt ctggaagcca agggctacaa agaagtgaaa aaggacctga tcatcaagct
     6961 gcctaagtac tccctgttcg agctggaaaa cggccggaag agaatgctgg cctctgccgg
     7021 cgaactgcag aagggaaacg aactggccct gccctccaaa tatgtgaact tcctgtacct
     7081 ggccagccac tatgagaagc tgaagggctc ccccgaggat aatgagcaga aacagctgtt
     7141 tgtggaacag cacaagcact acctggacga gatcatcgag cagatcagcg agttctccaa
     7201 gagagtgatc ctggccgacg ctaatctgga caaagtgctg tccgcctaca acaagcaccg
     7261 ggataagccc atcagagagc aggccgagaa tatcatccac ctgtttaccc tgaccaatct
     7321 gggagcccct gccgccttca agtactttga caccaccatc gaccggaaga ggtacaccag
     7381 caccaaagag gtgctggacg ccaccctgat ccaccagagc atcaccggcc tgtacgagac
     7441 acggatcgac ctgtctcagc tgggaggcga caagcgacct gccgccacaa agaaggctgg
     7501 acaggctaag aagaagaaag attacaaaga cgatgacgat aagggatccg gcgcaacaaa
     7561 cttctctctg ctgaaacaag ccggagatgt cgaagagaat cctggaccga ccgagtacaa
     7621 gcccacggtg cgcctcgcca cccgcgacga cgtccccagg gccgtacgca ccctcgccgc
     7681 cgcgttcgcc gactaccccg ccacgcgcca caccgtcgat ccggaccgcc acatcgagcg
     7741 ggtcaccgag ctgcaagaac tcttcctcac gcgcgtcggg ctcgacatcg gcaaggtgtg
     7801 ggtcgcggac gacggcgccg cggtggcggt ctggaccacg ccggagagcg tcgaagcggg
     7861 ggcggtgttc gccgagatcg gcccgcgcat ggccgagttg agcggttccc ggctggccgc
     7921 gcagcaacag atggaaggcc tcctggcgcc gcaccggccc aaggagcccg cgtggttcct
     7981 ggccaccgtc ggagtctcgc ccgaccacca gggcaagggt ctgggcagcg ccgtcgtgct
     8041 ccccggagtg gaggcggccg agcgcgccgg ggtgcccgcc ttcctggaga cctccgcgcc
     8101 ccgcaacctc cccttctacg agcggctcgg cttcaccgtc accgccgacg tcgaggtgcc
     8161 cgaaggaccg cgcacctggt gcatgacccg caagcccggt gcctgaacgc gttaagtcga
     8221 caatcaacct ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc
     8281 tccttttacg ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg
     8341 tatggctttc attttctcct ccttgtataa atcctggttg ctgtctcttt atgaggagtt
     8401 gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg caacccccac
     8461 tggttggggc attgccacca cctgtcagct cctttccggg actttcgctt tccccctccc
     8521 tattgccacg gcggaactca tcgccgcctg ccttgcccgc tgctggacag gggctcggct
     8581 gttgggcact gacaattccg tggtgttgtc ggggaaatca tcgtcctttc cttggctgct
     8641 cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct
     8701 caatccagcg gaccttcctt cccgcggcct gctgccggct ctgcggcctc ttccgcgtct
     8761 tcgccttcgc cctcagacga gtcggatctc cctttgggcc gcctccccgc gtcgacttta
     8821 agaccaatga cttacaaggc agctgtagat cttagccact ttttaaaaga aaagggggga
     8881 ctggaagggc taattcactc ccaacgaaga caagatctgc tttttgcttg tactgggtct
     8941 ctctggttag accagatctg agcctgggag ctctctggct aactagggaa cccactgctt
     9001 aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac
     9061 tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagggcc
     9121 cgtttaaacc cgctgatcag cctcgactgt gccttctagt tgccagccat ctgttgtttg
     9181 cccctccccc gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata
     9241 aaatgaggaa attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt
     9301 ggggcaggac agcaaggggg aggattggga agacaatagc aggcatgctg gggatgcggt
     9361 gggctctatg gcttctgagg cggaaagaac cagctggggc tctagggggt atccccacgc
     9421 gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac
     9481 acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt
     9541 cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc
     9601 tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc
     9661 gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact
     9721 cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg
     9781 gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc
     9841 gaattaattc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca
     9901 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca
     9961 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc
    10021 ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc
    10081 catggctgac taattttttt tatttatgca gaggccgagg ccgcctctgc ctctgagcta
    10141 ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctcccggga
    10201 gcttgtatat ccattttcgg atctgatcag cacgtgttga caattaatca tcggcatagt
    10261 atatcggcat agtataatac gacaaggtga ggaactaaac catggccaag ttgaccagtg
    10321 ccgttccggt gctcaccgcg cgcgacgtcg ccggagcggt cgagttctgg accgaccggc
    10381 tcgggttctc ccgggacttc gtggaggacg acttcgccgg tgtggtccgg gacgacgtga
    10441 ccctgttcat cagcgcggtc caggaccagg tggtgccgga caacaccctg gcctgggtgt
    10501 gggtgcgcgg cctggacgag ctgtacgccg agtggtcgga ggtcgtgtcc acgaacttcc
    10561 gggacgcctc cgggccggcc atgaccgaga tcggcgagca gccgtggggg cgggagttcg
    10621 ccctgcgcga cccggccggc aactgcgtgc acttcgtggc cgaggagcag gactgacacg
    10681 tgctacgaga tttcgattcc accgccgcct tctatgaaag gttgggcttc ggaatcgttt
    10741 tccgggacgc cggctggatg atcctccagc gcggggatct catgctggag ttcttcgccc
    10801 accccaactt gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt
    10861 tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg
    10921 tatcttatca tgtctgtata ccgtcgacct ctagctagag cttggcgtaa tcatggtcat
    10981 agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa
    11041 gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc
    11101 gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc
    11161 aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact
    11221 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac
    11281 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
    11341 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
    11401 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
    11461 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
    11521 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
    11581 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
    11641 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
    11701 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
    11761 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa
    11821 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
    11881 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
    11941 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
    12001 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
    12061 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
    12121 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
    12181 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
    12241 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag
    12301 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
    12361 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
    12421 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt
    12481 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
    12541 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg
    12601 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
    12661 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
    12721 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca
    12781 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct
    12841 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
    12901 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
    12961 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caa
//
